Transcriptional profiling of epidermal keratinocytes: Comparison of genes expressed in skin, cultured keratinocytes, and reconstituted epidermis, using large DNA microarrays
Transcriptional profiling of epidermal keratinocytes: Comparison of genes expressed in skin, cultured keratinocytes, and reconstituted epidermis, using large DNA microarrays
复制标题
DOI：
10.1111/j.1523-1747.2003.12611.x
复制
发表时间：
2003-12-01
影响因子：
6.5
通讯作者：
Blumenberg, M
中科院分区：
文献类型：
--
作者：
Gazel, A;Ramphal, P;Blumenberg, M
表皮角质形成细胞是复杂的细胞，其产生独特的三维（3-D）结构，通过多阶段过程分化，并对来自附近细胞的细胞外刺激做出反应。因此，角质形成细胞表达许多基因，即，具有相对较大的“转录组”。“为了确定哪些表达的基因是角质形成细胞固有的，哪些是分化和3-D结构特异性的，哪些是由其他细胞类型诱导的，我们比较了人类受试者皮肤的转录组，区分3-D重建表皮，培养的角质形成细胞和非角质形成细胞类型。使用大型寡核苷酸微阵列，我们分析了每种基因的五个或更多重复，产生了统计学一致的数据，并可以识别差异表达的基因。与其他细胞不同，表皮角质形成细胞表达许多蛋白酶和蛋白酶抑制剂以及保护免受紫外线照射的基因。皮肤特异性表达更多的受体、分泌蛋白和转录因子，这可能受到非角质形成细胞类型的影响。令人惊讶的是，皮肤中的线粒体蛋白质被显著抑制，表明代谢率低。三维样品，皮肤和重建的表皮，彼此相似，表达表皮分化标志物。培养的角质形成细胞表达许多细胞周期和DNA复制基因，以及整合素和细胞外基质蛋白。这些结果定义了表皮中的先天性、结构特异性和细胞类型调节基因。
Epidermal keratinocytes are complex cells that create a unique three-dimensional (3-D) structure, differentiate through a multistage process, and respond to extracellular stimuli from nearby cells. Consequently, keratinocytes express many genes, i.e., have a relatively large "transcriptome." To determine which of the expressed genes are innate to keratinocytes, which are specific for the differentiation and 3-D architecture, and which are induced by other cell types, we compared the transcriptomes of skin from human subjects, differentiating 3-D reconstituted epidermis, cultured keratinocytes, and nonkeratinocyte cell types. Using large oligonucleotide microarrays, we analyzed five or more replicates of each, which yielded statistically consistent data and allowed identification of the differentially expressed genes. Epidermal keratinocytes, unlike other cells, express many proteases and protease inhibitors and genes that protect from UV light. Skin specifically expresses a higher number of receptors, secreted proteins, and transcription factors, perhaps influenced by the presence of nonkeratinocyte cell types. Surprisingly, mitochondrial proteins were significantly suppressed in skin, suggesting a low metabolic rate. Three-dimensional samples, skin and reconstituted epidermis, are similar to each other, expressing epidermal differentiation markers. Cultured keratinocytes express many cell-cycle and DNA replication genes, as well as integrins and extracellular matrix proteins. These results define innate, architecture-specific, and cell-type-regulated genes in epidermis.